Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish

The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins, and which exert non-overlapping functions. However, there is little information about ontogenetically distinct neutrophil populations. In this work, using a photoconvertible transgenic zebrafish line; Tg(mpx:Dendra2), we selectively label rostral blood island-derived and caudal hematopoietic tissue-derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single-cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury, both populations share a core transcriptional profile as well as subset-specific transcriptional signatures. Accordingly, both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly, caudal neutrophils respond heterogeneously. Collectively, our results reveal that co-existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI-derived neutrophils in the head two times, at 34hpf and 44hpf, and performed caudal fin transections at 54hpf on the half of each group. Three hours later, we homogenized non-cut and cut embryos from each group and separately sorted red and green-fluorescent cells. The resulting four groups were analized by scRNAseq.